TSP-1、PDGF-B在早期糖尿病大鼠视网膜上的表达变化及曲安奈德的干预作用

发布时间：2018-01-16 21:12

本文关键词：TSP-1、PDGF-B在早期糖尿病大鼠视网膜上的表达变化及曲安奈德的干预作用　出处：《郑州大学》2011年硕士论文　论文类型：学位论文

【摘要】：糖尿病视网膜病变(DR)是糖尿病患者的严重并发症之一,是导致失明的重要原因,但DR的发病机制仍在探讨中。从DR的成因血流动态异常看,持续的高血糖状态、异常的血小板聚集和凝集不仅是引起毛细血管闭塞的重要因素,而且是促成视网膜缺血缺氧和眼内新生血管形成的主要病理生理基础。PDGF-B与TSP-1均由血小板释放,TSP-1已被公认为是一种有效的内源性血管生成抑制因子,PDGF-B广泛存在于发育中的血管,能够促进微血管周细胞生长,保持微血管的正常功能。近年来,国内(外)在DR成因中有关血流动力学方面关于血小板在DR晚期的研究比较多,而对早期的研究相对比较少。相关的研究表明,TSP-1、PDGF-B与新生血管的形成密切相关。曲安奈德是一种糖皮质激素,近些年来,广泛应用于眼科,但其对早期DR的治疗作用研究相对不多。本实验拟建立糖尿病眼动物模型,观察早期糖尿病大鼠视网膜上TSP-1(血小板反应蛋白-1)、PDGF-B (血小板源性生长因子-B)的表达变化及其相互关系,并探讨曲安奈德玻璃体腔注射在不同时期对其变化的干预作用。材料与方法选7周龄、健康、清洁级Wistar大鼠40只,体重150-160g,排除先天性近视及其它眼疾,随机分两组,每组20只,共40只,标准饲料饲养适应1周后,STZ诱导建立糖尿病大鼠模型。成模后,每周测血糖1次,将中间死亡大鼠及血糖恢复鼠弃除。其中Ⅰ组成模饲养4周后,实验眼玻璃体腔内注射曲安奈德液5ul(IA),对侧眼注射生理盐水5u1作为对照(IB),观察1周；Ⅱ组成模饲养6周后,实验眼玻璃体腔内注射曲安奈德液5ul (ⅡA),对侧眼注射生理盐水5ul作为对照(ⅡB),也观察1周；在此过程中感染眼内炎者大鼠均弃除,实验结束后,分别处死Ⅰ组和Ⅱ组大鼠,取眼球行视网膜切片HE染色、免疫组织化学染色,通过与电脑相连的Biosens Digital Imaging system分析系统对图像中阳性反应部位进行阳性平均灰度值分析,用阳性平均灰度值来表示免疫反应表达程度。阳性平均灰度值的高低与阳性表达的强弱成反比。结果免疫组化染色 TSP-1在视网膜上的表达：阳性细胞表现为胞浆着色呈棕黄色或黄色,主要位于视网膜神经节细胞层和内核层。阳性平均灰度值比较：ⅡA组与ⅠA组、ⅡA组与ⅡB组相比明显减少,差异有统计学意义(P0.05)；ⅡB与ⅠB组、ⅠA组与IB组相比无统计学意义。 PDGF-B在视网膜上的表达：阳性细胞表现为胞浆着色呈棕黄色或黄色,主要位于视网膜神经节细胞层和内核层。PDGF-B阳性平均灰度值比较：ⅠA组与ⅠB组、ⅡA组与ⅡB组相比明显减少,差异有统计学意义(P0.05)；ⅡA组与ⅠA组、ⅡB与ⅠB组相比无统计学意义。结论 1.STZ诱导成功建立糖尿病大鼠模型； 2.在早期DR阶段,曲安奈德应用能影响TSP-1在DR上的表达,推测TSP-1在早期即参与影响DR的发生； 3.在早期阶段,曲安奈德治疗能促进TSP-1和PDGF-B在糖尿病视网膜上的表达,推测在早期DR阶段,应用曲安奈德有助于DR的治疗。
[Abstract]:Diabetic retinopathy (DR) is one of the major complications of diabetes, is an important cause of blindness, but the pathogenesis of DR is still under discussion. See from the causes of DR abnormal blood flow, high blood sugar state, abnormal platelet aggregation and aggregation is not only the important factors caused by capillary occlusion, and is the main the pathophysiological basis of.PDGF-B and TSP-1 to retinal ischemia and intraocular neovascularization were released by platelets, TSP-1 has been recognized as a potent endogenous angiogenesis inhibitor, PDGF-B widely exists in the development of blood vessels, can promote the growth of microvascular pericytes, maintain the normal function of the microvascular in recent years. And domestic (foreign) in the DR formation in the relevant hemodynamic study on platelet in the late DR more, but the early research related research is relatively small. It indicates that TSP-1 and PDGF-B are closely related to the formation of neovascularization. Cu Ann Ned is a glucocorticoid. It has been widely applied in ophthalmology in recent years, but its research on early DR is relatively limited.
This experiment intends to establish the animal model of diabetic eye, observe the early diabetic rat retina TSP-1 (thrombospondin -1), PDGF-B (platelet-derived growth factor -B) expression and their relationship, and the effects of intravitreal injection of triamcinolone acetonide on changes in different period.
Materials and methods
7 weeks of age, health, clean Wistar rats 40, weight 150-160g, excluding congenital myopia and other eye diseases, were randomly divided into two groups, 20 rats in each group, a total of 40, the standard diet for 1 week after adaptation, the establishment of STZ induced diabetic rat model. After modeling, blood glucose was measured 1 times a week the death of rats, and rats. The blood glucose recovery discard compositions of die after 4 weeks of feeding, 5ul liquid injection of triamcinolone acetonide in the vitreous cavity (IA), contralateral injection of 5u1 physiologic saline as control (IB), 1 weeks of observation; II composition die after 6 weeks of feeding, 5ul liquid injection of triamcinolone acetonide the vitreous cavity (A II), contralateral injection of 5ul physiologic saline as control (B), and observed for 1 weeks; those rats were abandoned in infected endophthalmitis during this process, after the end of the experiment, respectively. Group I and group II rats, take the eye retinal slices stained with HE, immune histochemical staining with computer The Biosens Digital Imaging system analysis system was used to analyze the positive mean gray value of the positive part in the image, and the positive average gray value was used to indicate the expression level of the immune reaction. The positive mean gray level value was inversely proportional to the strength of the positive expression.
Result
Immunohistochemical staining
The expression of TSP-1 in the retina: the positive cells showed cytoplasmic staining was brown or yellow, mainly located in the retinal ganglion cell layer and the kernel layer. The positive average gray value comparison: A group and A group I, II, II A group decreased significantly compared with B group, the difference was statistically significant (P0.05); II I B and I B group, A group had no statistical significance compared with the IB group.
The expression of PDGF-B in the retina: the positive cells showed cytoplasmic staining was brown or yellow, mainly located in the retinal ganglion cell layer and core layer.PDGF-B positive average gray value comparison: A group and B group I I, II A group decreased significantly compared with B group, the difference was statistically significant (P0.05); II A group and A group 1, there was no significant difference between B and B II group.
conclusion
The diabetic rat model was successfully established by 1.STZ.
2. in the early DR stage, Cu Ann Ned application could affect the expression of TSP-1 on DR, and it was suggested that TSP-1 was involved in the occurrence of DR in the early stage.
3., in the early stage, Cu Ann Ned treatment can promote the expression of TSP-1 and PDGF-B on the diabetic retina. It is speculated that in the early stage of DR, the application of Cu Ann Ned is helpful for the treatment of DR.

【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R774.1

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